Kinetic parameters derived from Biacore analysis of interactions between BMP9, BMP10, and hAlk1-FC and mAlk1-FC

Kinetic analysis of receptor-ligand interactions studied by Biacore

Protein

hBMP9 immobilized on the chip

hBMP10 immobilized on the chip

kon (l/Ms)

koff(1/s)

KD (nmol/L)

kon (1/Ms)

koff (l/s)

KD (nmol/L)

hALK1-Fc

4.0 × 104

7.8 × 10−5

2.0

2.6 × 104

8.5 × 10−5

3.2

mALK1-Fc

3.5 × 104

9.4 × 10−5

2.3

2.3 × 104

1.1 × 10−4

4.7

Protein

hALK1-Fc immobilized on the chip

mALK1-Fc immobilized on the chip

BMP9

5.0 × 105

2.97 × 10−4

0.57

6.12 × 105

1.75 × 10−4

0.29

NOTE: The experiment was done on Biacore 3000 and data were processed with BIAevaluation software version 4.1. Ligands (BMP9 or BMP10) or receptors (hAlk1-Fc or mAlk1-Fc) were immobilized at low density by primary amine coupling on the CM5 chip. Different concentrations of analytes were injected at 30 μL/min over immobilized components. The best fit was obtained by using 1:1 binding model. The KD is calculated as koff/kon.